Pulp Revascularization in an Autotransplanted Mature Tooth: Visualization with Magnetic Resonance Imaging and Histopathologic Correlation.

Autotransplantation of a mature tooth usually leads to pulpal necrosis. Root canal treatment is recommended to prevent related inflammatory complications a few weeks after surgery. Extraoral root-end resection may facilitate reperfusion and obviate root canal treatment, but cannot be pictured with conventional dental radiography at this point in time. In the case of a lower mature transplanted molar, contrast-enhanced magnetic resonance imaging proved to be a feasible method for visualizing pulp revascularization just 4 weeks after autotransplantation. Consequently, root canal treatment was obviated. Nevertheless, the tooth had to be extracted 18 months postoperatively due to external cervical root resorption, probably caused by the extraction trauma. This allowed the histological processing and examination of the newly generated intracanal tissue. Uninflamed fibrovascular connective tissue was found, while odontoblasts or cementoblast-like cells were absent. These findings indicated that it was most likely stem cells from the bone marrow and the periodontal ligament that drove the regeneration.

Clinical, radiographic, and histological/histomorphometric analysis of maxillary sinus grafting with deproteinized porcine or bovine bone mineral: A randomized clinical trial.

OBJECTIVE: The present study aimed to compare histomorphometrically evaluated new bone formation, radiographically measured graft stability, and clinical implant outcome between maxillary sinus grafting with either deproteinized porcine bone mineral (DPBM) or deproteinized bovine bone mineral (DBBM). MATERIALS AND METHODS: Thirty maxillary sinuses were initially included and randomly assigned to the test group (TG; DPBM, n = 15) or control group (CG; DBBM, n = 15). After a healing period (6 months), axially retrieved bone biopsies of the molar region were used for histological/histomorphometric analysis of new bone formations. Additionally, radiographically measured graft stability and clinical implant outcome were assessed. RESULTS: Twenty-three sinus sites with 10 sinuses of the TG and 13 of the CG were ultimately available for data and statistical analysis. In the TG, a slightly, but yet significantly (p = .040) higher proportion of new bone formation (TG: 27.7 ± 5.6% vs. CG: 22.9 ± 5.1%) and a lesser (p = .019) amount of connective (non-mineralized) tissue (TG: 47.5 ± 9.5% vs. CG: 56.1 ± 9.5%) was found than in the CG. However, both xenografts showed comparable (n.s.) residual bone graft (TG: 23.7 ± 7.2% vs. CG: 21.1 ± 9.85.6%), bone-to-graft contacts (TG: 26.2 ± 9.8% vs. CG: 30.8 ± 13.8%), similar graft height reduction over time (TG: 12.9 ± 6.7% CG: 12.4 ± 5.8%) and implant survival/success rate (100%). At the 3-year post-loading evaluation, the peri-implant marginal bone loss (TG: 0.52 ± 0.19 mm; CG: 0.48 ± 0.15 mm) and the peri-implant health conditions (TG: 87.5%/CG: 81.2%) did not differ between implants inserted in both xenografts used. CONCLUSIONS: The use of DPBM or DBBM for maxillary sinus augmentation is associated with comparable bone formation providing stable graft dimension combined with healthy peri-implant conditions.

Mg-Zn-Ca Alloy (ZX00) Screws Are Resorbed at a Mean of 2.5 Years After Medial Malleolar Fracture Fixation: Follow-up of a First-in-humans Application and Insights From a Sheep Model.

BACKGROUND: In the ongoing development of bioresorbable implants, there has been a particular focus on magnesium (Mg)-based alloys. Several Mg alloys have shown promising properties, including a lean, bioresorbable magnesium-zinc-calcium (Mg-Zn-Ca) alloy designated as ZX00. To our knowledge, this is the first clinically tested Mg-based alloy free from rare-earth elements or other elements. Its use in medial malleolar fractures has allowed for bone healing without requiring surgical removal. It is thus of interest to assess the resorption behavior of this novel bioresorbable implant. QUESTIONS/PURPOSES: (1) What is the behavior of implanted Mg-alloy (ZX00) screws in terms of resorption (implant volume, implant surface, and gas volume) and bone response (histologic evaluation) in a sheep model after 13 months and 25 months? (2) What are the radiographic changes and clinical outcomes, including patient-reported outcome measures, at a mean of 2.5 years after Mg-alloy (ZX00) screw fixation in patients with medial malleolar fractures? METHODS: A sheep model was used to assess 18 Mg-alloy (ZX00) different-length screws (29 mm, 24 mm, and 16 mm) implanted in the tibiae and compared with six titanium-alloy screws. Micro-CT was performed at 13 and 25 months to quantify the implant volume, implant surface, and gas volume at the implant sites, as well as histology at both timepoints. Between July 2018 and October 2019, we treated 20 patients with ZX00 screws for medial malleolar fractures in a first-in-humans study. We considered isolated, bimalleolar, or trimalleolar fractures potentially eligible. Thus, 20 patients were eligible for follow-up. However, 5% (one patient) of patients were excluded from the analysis because of an unplanned surgery for a pre-existing osteochondral lesion of the talus performed 17 months after ZX00 implantation. Additionally, another 5% (one patient) of patients were lost before reaching the minimum study follow-up period. Our required minimum follow-up period was 18 months to ensure sufficient time to observe the outcomes of interest. At this timepoint, 10% (two patients) of patients were either missing or lost to follow-up. The follow-up time was a mean of 2.5 ± 0.6 years and a median of 2.4 years (range 18 to 43 months). RESULTS: In this sheep model, after 13 months, the 29-mm screws (initial volume: 198 ± 1 mm3) degraded by 41% (116 ± 6 mm3, mean difference 82 [95% CI 71 to 92]; p < 0.001), and after 25 months by 65% (69 ± 7 mm3, mean difference 130 [95% CI 117 to 142]; p < 0.001). After 13 months, the 24-mm screws (initial volume: 174 ± 0.2 mm3) degraded by 51% (86 ± 21 mm3, mean difference 88 [95% CI 52 to 123]; p = 0.004), and after 25 months by 72% (49 ± 25 mm3, mean difference 125 [95% CI 83 to 167]; p = 0.003). After 13 months, the 16-mm screws (initial volume: 112 ± 5 mm3) degraded by 57% (49 ± 8 mm3, mean difference 63 [95% CI 50 to 76]; p < 0.001), and after 25 months by 61% (45 ± 10 mm3, mean difference 67 [95% CI 52 to 82]; p < 0.001). Histologic evaluation qualitatively showed ongoing resorption with new bone formation closely connected to the resorbing screw without an inflammatory reaction. In patients treated with Mg-alloy screws after a mean of 2.5 years, the implants were radiographically not visible in 17 of 18 patients and the bone had homogenous texture in 15 of 18 patients. No clinical or patient-reported complications were observed. CONCLUSION: In this sheep model, Mg-alloy (ZX00) screws showed a resorption to one-third of the original volume after 25 months, without eliciting adverse immunologic reactions, supporting biocompatibility during this period. Mg-alloy (ZX00) implants were not detectable on radiographs after a mean of 2.5 years, suggesting full resorption, but further studies are needed to assess environmental changes regarding bone quality at the implantation site after implant resorption. CLINICAL RELEVANCE: The study demonstrated successful healing of medial malleolar fractures using bioresorbable Mg-alloy screws without clinical complications or revision surgery, resulting in pain-free ankle function after 2.5 years. Future prospective studies with larger samples and extended follow-up periods are necessary to comprehensively assess the long-term effectiveness and safety of ZX00 screws, including an exploration of limitations when there is altered bone integrity, such as in those with osteoporosis. Additional use of advanced imaging techniques, such as high-resolution CT, can enhance evaluation accuracy.

Degradation behavior and osseointegration of Mg-Zn-Ca screws in different bone regions of growing sheep: a pilot study.

Magnesium (Mg)-based implants are highly attractive for the orthopedic field and may replace titanium (Ti) as support for fracture healing. To determine the implant-bone interaction in different bony regions, we implanted Mg-based alloy ZX00 (Mg < 0.5 Zn < 0.5 Ca, in wt%) and Ti-screws into the distal epiphysis and distal metaphysis of sheep tibiae. The implant degradation and osseointegration were assessed in vivo and ex vivo after 4, 6 and 12 weeks, using a combination of clinical computed tomography, medium-resolution micro computed tomography (µCT) and high-resolution synchrotron radiation µCT (SRµCT). Implant volume loss, gas formation and bone growth were evaluated for both implantation sites and each bone region independently. Additionally, histological analysis of bone growth was performed on embedded hard-tissue samples. We demonstrate that in all cases, the degradation rate of ZX00-implants ranges between 0.23 and 0.75 mm/year. The highest degradation rates were found in the epiphysis. Bone-to-implant contact varied between the time points and bone types for both materials. Mostly, bone-volume-to-total-volume was higher around Ti-implants. However, we found an increased cortical thickness around the ZX00-screws when compared with the Ti-screws. Our results showed the suitability of ZX00-screws for implantation into the distal meta- and epiphysis.

Size changes in miR­21 knockout mice: Geometric morphometrics on teeth, alveolar bone and mandible.

MicroRNA­21 (miR­21) is a small non­coding RNA that is differentially expressed during tooth development, particularly during amelogenesis. Although orthodontic tooth movement and the innate immune response are impaired, miR­21 knockout mice demonstrate no obvious skeletal phenotype. However, the consequence of miR­21 knockout on tooth phenotype and corresponding alveolar bone is unknown. The current study utilized landmark­based geometric morphometrics to identify anatomical dissimilarities of the three lower and upper molars, and the corresponding alveolar bone, in miR­21 knockout and wild­type control mice. The anatomical structures were visualized by microcomputer tomography. A total of 36 and 38 landmarks were placed on mandibular and maxillary molars, respectively. For the alveolar bone, 16 landmarks were selected on both anatomical sites. General Procrustes analysis revealed significantly smaller molars and dimensions of the alveolar bone in the mandible of the miR­21 knockout mice when compared with wild­type controls (P=0.03 and P=0.04, respectively). The overall dimension of the mandible was reduced by the lack of miR­21 (P=0.02). In the maxilla, the dimension of the alveolar bone was significant (P=0.02); however, this was not observed in the molars (P=0.36). Based on principal component analysis, no changes in shape for any of the anatomical sites were observed. Dental and skeletal jaw length were calculated and no prognathism was identified. However, the fluctuating asymmetry of the molars in the mandible and the maxilla was reduced in the miR­21 knockout mice by 38 and 27%, respectively. Taken together, the results of the present study revealed that the molars in the mandible and the dimension of the respective alveolar bone were smaller in miR­21 mice compared with wild­type littermates, suggesting that miR­21 influences tooth development.

Caspase inhibitor attenuates the shape changes in the alveolar ridge following tooth extraction: A pilot study in rats.

OBJECTIVE: The aim of the study was to determine whether the inhibition of apoptosis via pan-caspase inhibitors can attenuate the changes in the alveolar ridge upon tooth extraction. BACKGROUND: Cells undergoing apoptosis might play a central role in the onset of alveolar bone resorption and the ensuing bone atrophy following tooth extraction. Caspases are proteases that regulate apoptotic cell death. It is, therefore, reasonable to hypothesize that blocking apoptosis with pan-caspase inhibitors attenuates the changes in the alveolar ridge following tooth extraction. METHODS: In 16 inbred rats, the mandibular first (M1) and second (M2) molars of one side were extracted. Following random allocation, the rats received either a cell-permeable pan-caspase inhibitor or diluent. After a healing period of 10 days, changes in shape and height of the alveolar ridge were examined using geometric morphometrics and linear measurements based on micro-computed tomography. RESULTS: Geometric morphometric analysis revealed that the pan-caspase inhibitor prevented major shape changes of the alveolar ridge following M1 tooth extraction (P < .05). Furthermore, linear measurements confirmed that the pan-caspase inhibitor significantly prevented the atrophy of the alveolar ridge height following M1 tooth extraction compared to the diluent controls (-0.53 mm vs -0.24 mm; P = .012). M2 tooth extraction caused no shape changes of the alveolar ridge, and thus, the pan-caspase inhibitor group did not differ from the control group (-0.14 mm vs -0.05 mm; P = .931). CONCLUSIONS: These findings suggest that the inhibition of apoptosis may attenuate shape changes of the alveolar ridge following M1 tooth extraction in rodents.

ATG7 is essential for secretion of iron from ameloblasts and normal growth of murine incisors during aging.

The incisors of rodents comprise an iron-rich enamel and grow throughout adult life, making them unique models of iron metabolism and tissue homeostasis during aging. Here, we deleted Atg7 (autophagy related 7) in murine ameloblasts, i.e. the epithelial cells that produce enamel. The absence of ATG7 blocked the transport of iron from ameloblasts into the maturing enamel, leading to a white instead of yellow surface of maxillary incisors. In aging mice, lack of ATG7 was associated with the growth of ectopic incisors inside severely deformed primordial incisors. These results suggest that 2 characteristic features of rodent incisors, i.e. deposition of iron on the enamel surface and stable growth during aging, depend on autophagic activity in ameloblasts. Abbreviations: ATG5: autophagy related 5; ATG7: autophagy related 7; CMV: cytomegalovirus; Cre: Cre recombinase; CT: computed tomography; FTH1: ferritin heavy polypeptide 1; GFP: green fluorescent protein; KRT5: keratin 5; KRT14: keratin 14; LGALS3: lectin, galactose binding, soluble 3; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; NCOA4: nuclear receptor coactivator 4; NRF2: nuclear factor, erythroid 2 like 2; SQSTM1: sequestosome 1.

Osteointegration of a Novel Silk Fiber-Based ACL Scaffold by Formation of a Ligament-Bone Interface.

BACKGROUND: Given the unsatisfactory results and reported drawbacks of anterior cruciate ligament (ACL) reconstruction, such as donor site morbidity and the limited choice of grafts in revision surgery, new regenerative approaches based on tissue-engineering strategies are currently under investigation. PURPOSES: To determine (1) if a novel silk fiber-based ACL scaffold is able to initiate osteointegration in the femoral and tibial bone tunnels under in vivo conditions and (2) if the osteointegration process will be improved by intraoperatively seeding the scaffolds with the autologous stromal vascular fraction, an adipose-derived, stem cell-rich isolate from knee fat pads. STUDY DESIGN: Controlled laboratory study. METHODS: A total of 33 sheep underwent ACL resection and were then randomly assigned to 2 experimental groups: ACL reconstruction with a scaffold alone and ACL reconstruction with a cell-seeded scaffold. Half of the sheep in each group were randomly chosen and euthanized 6 months after surgery and the other half at 12 months. To analyze the integration of the silk-based scaffold in the femoral and tibial bone tunnels, hard tissue histology and micro-computed tomography measurements were performed. RESULTS: Hard tissue histological workup showed that in all treatment groups, with or without the application of the autologous stromal vascular fraction, an interzone of collagen fibers had formed between bone and silk-based graft. This collagen-fiber continuity partly consisted of Sharpey fibers, comparable with tendon-bone healing known for autografts and allografts. Insertion sites were more broad based at 6 months and more concentrated on the slightly protruding, bony knoblike structures at 12 months. Histologically, no differences between the treatment groups were detectable. Analysis of micro-computed tomography measurements revealed a significantly higher tissue density for the cell-seeded scaffold group as compared with the scaffold-alone group in the tibial but not femoral bone tunnel after 12 months of implantation. CONCLUSION: The novel silk fiber-based scaffold for ACL regeneration demonstrated integration into the bone tunnels via the formation of a fibrous interzone similar to allografts and autografts. Histologically, additional cell seeding did not enhance osteointegration. No significant differences between 6 and 12 months could be detected. After 12 months, there was still a considerable amount of silk present, and a longer observation period is necessary to see if a true ligament-bone enthesis will be formed. CLINICAL RELEVANCE: ACL regeneration with a silk fiber-based scaffold with and without additional cell seeding may provide an alternative treatment option to current techniques of surgical reconstruction.

Cell culture models of oral mucosal barriers: A review with a focus on applications, culture conditions and barrier properties.

Understanding the function of oral mucosal epithelial barriers is essential for a plethora of research fields such as tumor biology, inflammation and infection diseases, microbiomics, pharmacology, drug delivery, dental and biomarker research. The barrier properties are comprised by a physical, a transport and a metabolic barrier, and all these barrier components play pivotal roles in the communication between saliva and blood. The sum of all epithelia of the oral cavity and salivary glands is defined as the blood-saliva barrier. The functionality of the barrier is regulated by its microenvironment and often altered during diseases. A huge array of cell culture models have been developed to mimic specific parts of the blood-saliva barrier, but no ultimate standard in vitro models have been established. This review provides a comprehensive overview about developed in vitro models of oral mucosal barriers, their applications, various cultivation protocols and corresponding barrier properties.

Comparison of marginal fit of cemented zirconia copings manufactured after digital impression with lava™ C.O.S and conventional impression technique.

BACKGROUND: Evaluation of the marginal fit of cemented zirconia copings manufactured after digital impression with Lava™ Chairside Oral Scanner in comparison to that of zirconia copings manufactured after conventional impressions with polyvinyl siloxane. METHODS: A prepared typodont tooth #36, was replicated 40 times with a vinyl silicone and precise model resin. The dies were randomly divided into two groups according to the impression taking technique. Digital impressions with Lava™ C.O.S. and conventional impressions were taken according to the group. Subsequently zirconia copings were manufactured and cemented on their respective dies with zinc oxide phosphate cement. After embedding in resin, mesio-distal section of each coping was performed with a diamond saw in order to obtain two slices. One half of the specimen was used for evaluation with an optical microscope (OM) and the other half for evaluation with a scanning electron microscope (SEM). Marginal gap (MG) and absolute marginal discrepancy (AMD) were measured mesial and distal on each slice. RESULTS: No significant difference of the marginal parameters between the digital and the conventional group was found. The mean values for MG in the digital group were 96.28 µm (+/-43.21 µm) measured with the OM and 99.26 µm (+/-48.73 µm) measured with the SEM, respectively. AMD mean values were 191.54 µm (+/-85.42 µm) measured with the optical microscope and 211.6 µm (+/-96.55 µm) with the SEM. For the conventional group the mean MG values were 94.84 µm (+/-50.77 µm) measured with the OM and 83.37 µm (+/-44.38 µm) measured with the SEM, respectively. AMD mean values were 158.60 µm (+/-69.14 µm) for the OM and 152.72 µm (+/-72.36) for the SEM. CONCLUSIONS: Copings manufactured after digital impression with Lava™ C.O.S. show comparable marginal parameters with the copings manufactured after conventional impression with polyvinyl syloxane. The mean MG values of both groups fit in the clinically acceptable range.

Alveolar bone regeneration in response to local application of calcitriol in vitamin D deficient rats.

AIM: Vitamin D deficiency is considered to diminish bone regeneration. Yet, raising the serum levels takes months. A topic application of the active vitamin D metabolite, calcitriol, may be an effective approach. Thus, it becomes important to know the effect of vitamin D deficiency and local application on alveolar bone regeneration. MATERIAL AND METHODS: Sixty rats were divided into three groups; two vitamin depletion groups and a control group. Identical single defects (2 mm diameter) were created in the maxilla and mandible treated with calcitriol soaked collagen in one deficiency group while in the other two groups not. Histomorphometric analysis and micro CTs were performed after 1 and 3 weeks. Serum levels of 25(OH)D3 and PTH were determined. RESULTS: Bone formation rate significantly increased within the observation period in all groups. Bone regeneration was higher in the maxilla than in the mandible. However, bone regeneration was lower in the control group compared to vitamin depletion groups, with no significant effects by local administration of calcitriol (micro CT mandible p = 0.003, maxilla p < 0.001; histomorphometry maxilla p = 0.035, mandible p = 0.18). CONCLUSION: Vitamin D deficiency not necessarily impairs bone regeneration in the rat jaw and a single local calcitriol application does not enhance healing.

A histomorphometric analysis of the nature of the mandibular canal in the anterior molar region.

OBJECTIVES: Knowledge of the position and configuration of the mandibular canal is a basic requirement before implant placement in the mandible. Radiological studies suggest a positive correlation between alveolar trabecular bone quality and mandibular canal corticalization. The aim of this study was to test this assumption histomorphometrically in the anterior molar region, which is one of the most frequent places for implantation. MATERIALS AND METHODS: Fifty thin ground sections (from 28 male and 22 female cadavers) of the first molar region were investigated for trabecular bone volume and thickness and the presence of a mandibular canal wall. RESULTS: Trabecular bone volume was significantly higher in males (p = 0.009). Further, it correlated significantly with the presence of a canal wall (rho = 0.585, p < 0.001), indicating that a reduced trabecular bone volume is associated with a reduced amount of bone surrounding the alveolar nerve. The cranial aspects of the canal wall were present at a significantly lower frequency (64.64 %) than the buccal, lingual, or caudal sides (p < 0.006). CONCLUSION: The present study demonstrated that low trabecular bone volume correlates with only a fragmentarily present mandibular canal wall. This suggests that bone surrounding the alveolar nerve is of trabecular, not cortical, origin and possibly affected by reduction of the trabecular bone. CLINICAL RELEVANCE: These results imply that oral surgeons should pay particular attention to implant placement in patients with low alveolar bone quality. The cranial aspects of the mandibular canal might be only fragmentarily or even completely missing. Consequently, they hardly present resistance during implant site preparation, and the risk for nerve injury, e.g., due to post-surgery hematoma, could be increased.